SE453092B - NEW CEPHALOSPORIN DERIVATIVES, PROCEDURES FOR PREPARING THEREOF AND COMPOSITIONS THEREOF - Google Patents

Description

453 092 generically a large number of cephalosporin derivatives of the form Ra N _ .t fi l / i == H .__ (LI u R 1- s \ oa2 Å / A coon 3 where the substituents R1, R2, R3, R4, X and A generically include the corresponding substituents in the compounds of formula I above, however, in the 20 column long definition of the various substituent groups, in the 78 page list of structural formulas and in the 225 working examples there is no disclosure that A could be a quaternary ammonium methyl group of the The British Patent Specification 1,604,971 corresponds to the said U.S. Patent Specification and has a substantially identical content.The published British Patent Application 2,028,305 A exhibits the same broad generic content, although it discloses is not formally related to the just mentioned patents, but A is exemplified only as hydrogen.

U.S. Patent 4,278,671 discloses 7- (2- (2-aminothiazol-4-yl) -2- (α] -methoxyiminoacetamido) cephalosporin derivatives of the formula SS ε-conz-xw / 9033 CH 2 RB OOH wherein R 2 NH is a optionally protecting amino group and R 3 is hydrogen or "the residue of a nucleophilic compound". The term "residue of a nucleophilic compound" encompasses a broad definition, indicating that R 3 is a quaternary ammonium group. "Only pyridinium, alternatively pyridinium, quinolinium, picolinium and lutidinium substituted can be disclosed as possible quaternary. There is no indication that the quaternary ammonium group may be of the type included in the compounds of formula I above. British Patent Specification 1,581,854 corresponds to the said U.S. Patent Specification and has a substantially identical content. on the same patent holders, which are not formally related to the aforementioned patent but which have similar content, include U.S. Patents 4,098,888, 4,203,899, 4,205,180 and 4,298,606 and British Patent Specification 1,536,281.

Published British Patent Specification 2,040,921 discloses cephalosporin derivatives of the formula NH2 AN K - ¿S 'C CONH 4_J / I 1 u =' n \ o-ï-coon är. / 329% * 32 uh coca å: wherein Ra and Rb are C 1-4 alkyl or together with the carbon atom to which they are attached form a C 3-7 cycloalkylidene ring and R 1, R 2 and R 3 are each a C 1-4 alkyl- 9fuPP- Presented invention relates to cephalosporin derivatives with uno s ®R3 N / cnz-n-cu In formula II 3 c | wherein R 1 is hydrogen, R 2 and R 3 are each independently methyl or ethyl and R 4 is methyl, ethyl, allyl, 2-hydroxyethyl, 2- (dimethylamino) ethyl or pyridylmethyl , and non-toxic pharmaceutically acceptable acid addition salts thereof as well as processes for their preparation. Also within the scope of the present invention are solvates (including hydrates) of the compounds of formula I as well as the tautomeric forms of the compounds of formula I, for example the 2-iminothiazolin-4-yl form of the 2-aminothiazol-4-yl group.

As shown in the structural formula above, the compounds of formula I have the "syn" or "Z" configuration with respect to the alkoxyimino group. Since the compounds are geometric isomers, some amount of the "anti" isomer may also be present. Formula I which contains at least 90% of the "syn" isomer.

Preferably, the compounds of formula I are "syn" isomers which are substantially free of the corresponding "anti" isomers.

The non-toxic, pharmaceutically acceptable acid addition salts of the compounds of formula I include the salts with hydrochloric acid, hydrobromic acid, formic acid, nitric acid, sulfuric acid, methanesulfonic acid, phosphoric acid, acetic acid and trifluoroacetic acid and other acids which have been used in penicillin. cephalosporin area.

The compounds of formula I in which R 1 is hydrogen show high antibacterial activity with respect to various Gram-positive and Gram-negative bacteria and are useful for the treatment of bacterial infections in animals including humans. The compounds of formula I may be formulated for parenteral use in a conventional manner using known pharmaceutical carriers and excipients and may be presented in unit dosage form or in multiple dosage form. The compositions may be in the form of solutions, suspensions or emulsions in oily or aqueous vehicles and may contain conventional dispersing, suspending or stabilizing agents. The compositions may also be in the form of a dry powder for reconstitution before use, for example with sterile pyrogen-free water. The compounds of formula I may also be prepared as suppositories using conventional suppository bases such as cocoa butter or other glycerides.

The compounds of the present invention may, if desired, be administered in combination with other antibiotics such as penicillins or other cephalosporins.

When formulated in unit dosage form, the compositions preferably contain 50 to 1500 mg of the active ingredient of formula I. The dosage of the compounds of formula I depends on such factors as the weight and age of the patient as well as the particular nature and severity of the disease. and is ultimately decided by the doctor.

However, the dosage for the treatment of an adult is in the range of 50-5000 mg per day depending on the frequency of administration and the mode of administration. For intramuscular or intravenous administration to adults, a total dose of 750-3000 mg per day, in divided doses, is usually sufficient, although higher daily doses of certain compounds may be desirable in the case of Pseudomonas infections.

The preferred compounds of formula I are those wherein R 1 is hydrogen, R 2 is methyl, R 3 is methyl or ethyl and R 4 is methyl, ethyl, 2-hydroxyethyl, 2- (dimethylamino) ethyl, allyl or pyridylmethyl. Particularly preferred compounds are those wherein R 1 is hydrogen, R 2 and R 3 are methyl and R 4 is methyl, 2-hydroxyethyl or allyl. The most preferred compound is that R 1 is hydrogen and R 2, R 3, R 4 are methyl. In the preliminary evaluation of the preferred compounds of the present invention, the minimum inhibitory concentrations (MIC) of the compounds were determined by the dual agar series dilution method in Mueller-Hinton agar with respect to 32 strains of the test organisms in 6 groups.

The geometric mean values of MIK data in this test are given in Table 1 below. 453 092 MH> fl .HUWQNW .NMHHO Now fm. .ëää m .. .mm "HHTTQ .šw N. .Mmm: oo .äêæuw N. mmwä .ME .Eßm S Wwnwmw fl om .Mm fi HTTø. .äšßm m. Jim: oo ... fi mëšm m. S8 .m» äu fi wwhv fi po fi mwwu u fi- Tø.

Aumšßm N. .mm fi o .aßw: .ü ~ .uëaßm N. 38 .m m fl mnwvv fi uo fl mwwu u wTTw.

.Hås fi w m. Määa .m »äa fi mwnà flfififi u fi æm n åiö. .äašw m. äwäm .m w fi ß fl fñ fififl u fiäm n wTfó.

Q ».Ä 36 Bo ... m6 .ä w i» 3.0 -6 Se ... ä .. få w w fi Sa Nwa S56 ... m w; m .Hm 85 mi .. mmo ... ä .. w; N ... ïïm mwåäws. QNSBN ... mwoazmmoa. wäæñm wii; E ..

S. E. G. E. S. Cšøšm m.

HHTTo. Eino. nTTo. Sludge. QTTo. mHnïw. . .ä .Em .Ešë ä: .då w fi äwš fi wwm .. »ä fifl ëöww maëmää _. The absorbance of the most preferred compound Ia (prepared according to Example 1) was determined in mice after a single intramuscular injection of the test compound (dissolved in 0.1 M phosphate buffer; pH 7) at a dosage of 20 mg / kg.

Blood samples were drawn from the eye sockets and collected in heparinized capillary tubes and analyzed in Mueller-Hinton medium using Morganella morganii A9695 as a test organism. The blood levels at different time intervals, the half-life values (t1 / 2) and the areas under the curve (YUK) are shown in Table 2 below.

Table 2 Blood levels of compound in mice Blood levels 10 14 pg / ml 20 12 at minutes 30 8.8 after 40 7.5 administration 50 4.7 60 4.4 90 1.5 120 0.74 * 1/2 24 (minutes ) YUK pg-hr / ml 1 10 453 092 The in vitro activity of the most preferred compound Ia with respect to 31 strains of severely treated bacteria was largely determined by GC agar and the results are shown in Table 3 below.

Table 3 In vitro activity of compound Ia with respect to severely treated bacteria Test organism (ampicillin-resistant) Geometric mean of MIK (pg / ml) §, Exogenesis (6 strains) 0.013 §, Eneumoniae (6) 0.013 _§. gonorrhoeae (4) 0.013 §. meningitidis (5) 0.016 ä ». (7) 0.013 (ampicillin-sensitive) fi. influenzae (3) 0,20,453,092 According to another aspect, the invention relates to processes for the preparation of the compounds of formula I. There are two main processes for the conversion of an readily available starting cephalosporin to an æumn cephalosporin with different substituents in the 7th and 3rd positions. One can first remove the 7-substituent and replace it with the desired 7-substituent and then introduce the desired 3-substituent.

Alternatively, one can first introduce the desired 3-substituent and then replace the 7-substituent. The compounds of formula I can be prepared by either method and both fall within the scope of the present invention, but it is preferred to introduce the desired 7-substituent first and then to introduce the desired 3-substituent. The preferred process is set forth in Reaction Scheme 1 below while the alternative process is set forth in Reaction Scheme 2 below. The abbreviation "Tr" represents the trityl group (triphenylmethyl), which is a preferred amino protecting group.

The abbreviation "Ph" represents the phenyl group. Thus, the -CH (Ph) 2 group is the benzhydryl group, which is a preferred carboxyl protecting group.

Reaction Scheme 1 N c-cooczns Ä \ §u H Trim S \ on azx C - C0OC2H5 Å u _ III TIHN S \\\ 0R2 ou '453 092 HWÄWÉOÃÛH r - N o / f / cazcl coocrxæmz V / 5 C CONH Ä 3 ”Ä f 'rrrm s \ 0R2 0% azel coocumüz NaI 1; \: á CONH l / s f> / L \ 2 o / y-N / H21 coocnmn) 2 RB / \ R4 2. deblocking 1. GRs-N VI VII 455 092 11 C CONR 3 Å å u \ 2 'r- * E' / å S OR O Clii- T: CH3 Reaktionssèhema 2 s Phen com; -T 2 7 vn: fn / azon COOCH (Ph) 2 PCIS pyridine s Phcnzcøuaw-W / IX O / l-'N / azel coocu (m) 2 NaI 453 092 12's Phcxzcouu- ß az: coocz fl vrnz / Ra ca -N 3 \\\ R4 hn Conn svc 2 på: / "'_“ / a - --c1-x of 2 3 coocmrmz R4 deacylering n x- s 2' 19 123 04 "” / ai-uf-caa l coocacvrn, n * XI XII 453 092 13 S °° “**" "" '"J / 33 xnr ._- Gl TrHN' S \ op_2 (f / HfZ-ï-C fl s cooczurn) 2 n 'avblockering xV Ehuru, The above reaction schemes show preferred multi-step processes for the preparation of the compounds of formula I it will be appreciated that other starting materials and processes may be used for the preparation of the intermediates used in the key step of each reaction scheme. Thus, the key step in Reaction Scheme 1 is the reaction of compound VII with the tertiary amine. Compound VII can be prepared by other methods itself. Similarly, the key step in Reaction Scheme 2 is the acylation of compound XII with compound IV. Both compounds XII and IV can be prepared by other methods.

The present invention provides a process for the preparation of compounds of the formula 453 092 14 s n wherein R 2 and R 3 are each independently methyl or ethyl and R 4 is methyl, ethyl, allyl, 2-hydroxyethyl, 2- (dimethylamino) ethyl or pyridylmethyl, and non-toxic pharmaceutically acceptable salts and solvates thereof, which means a compound having the reaction of a compound having the formula B1 is a conventional carboxyl protecting group B2 is a conventional amino protecting group having a tertiary amine of the formula / RB 4 CH3-N R wherein R3 and R4 have the meanings given above, for the preparation of a compound of the formula C008 fi; .__ m -1/8 u 'i 19 113 szm / aï fi \ onz o // r- \; L1cu2-§:% - cr R4 and that all protective groups are then removed in a conventional manner.

The reaction is carried out in an anhydrous organic solvent such as methylene chloride, chloroform, ethyl ether, hexane, ethyl acetate, tetrahydrofuran, acetonitrile and the like or mixtures of such solvents. The reaction is conveniently carried out at a temperature of from -10 ° C to + 50 ° C; normally it is preferable to carry out the reaction at room temperature. At least one mole of the tertiary amine should be used per mole of compound XIV; normally it is preferred to utilize 50-100% excess of the tertiary amine.

Carboxyl protecting groups suitable for use as B1 in the above reaction are well known to those skilled in the art and include aralkyl groups such as benzyl, p-methoxybenzyl, p-nitrobenzyl and diphenylmethyl (benzhydryl); alkyl groups such as p-butyl, haloalkyl groups such as 2,2,2-trichloroethyl and other carboxyl protecting groups described in the literature, for example in British Patent Specification 1 399 086. It is preferred to use carboxyl protecting groups which can be easily removed by treatment. with acid. Particularly preferred carboxyl protecting groups are the benzhydryl and t-butyl groups.

Amum protecting groups suitable for use as B2 are also well known in the art and include the triethyl group and acyl groups such as chloroacetyl. Amino protecting groups which can be easily removed by treatment with acid, for example the trityl group, are preferred.

The present invention also encompasses a process for the preparation of compounds of the formula ° s fn in "beer JH s \ ,, R2: V4 / caz-u-c fl a I 2 om? N 'wherein R2 and R3 are each independently methyl or ethyl and R 4 is methyl, ethyl, allyl, 2-hydroxyethyl, 2- (dimethyl-B KN N 2 - u 2 - m fi ç- (s: IQ Ra xv 2 \ ° R2 Go-N / lcuz-Qaï-cna 453 092 16 amino ) ethyl or pyridylmethyl, and non-toxic pharmaceutically acceptable salts and solvates thereof, which process acylates a compound with the compound “T fl (sp Rs rN /. ® | 0 / GHz- N- C143 1 | 4 R 1 XVI COOB or an N-silyl derivative thereof, wherein B 1 is hydrogen or a conventional carboxyl protecting group and R 3 and R 4 have the meanings given above, with an acylating derivative of an acid of the formula Bzrm / äsïrëïoíïz wherein B2 is a conventional amino protecting group and R XVII 2 is of the significance given above, for the preparation of a compound of the formula S C008 RAW and that all protective groups are subsequently removed.

The acylating derivatives of the acid of formula XVII include the acid halides (and especially the acid chloride), mixed acid anhydrides (such as the acid anhydrides formed with pivalic acid or a haloformate such as ethyl chloroformate) and activated esters) which may be formed with N-hydroxybenstriazole in the presence of a condensation dicyclohexylcarbodiimide). The acylation can also be carried out using the free acid of formula XVII in the presence of a condensing agent such as dicyclohexylcarbodiimide, carbonyldiimidazole or an isoxazolium salt. The term "acylating derivative" of the acid of formula XVII in the present context includes the free acid itself in the presence of a condensing agent as described above. The preferred acylating derivative of the acid of formula XV§I is the acid chloride which is preferably used in the presence of an acid-binding agent (and especially a tertiary amine as an acid-binding agent such as triethylamine, dimethylaniline or pyridine).

When the acylation is carried out with an acid halide, it is possible to use an aqueous reaction medium, but an anhydrous medium is preferred. When acid anhydrides, activated esters or the free acid in the presence of a condensing agent are used for the acylation, the reaction medium should be anhydrous. Particularly preferred solvents for the acylation reaction are halogenated hydrocarbons such as ty ßtyha fl d flfl ïd and chloroform, but tertiary amides such as dimethylacetamide or dimethylformamide may also be used as well as other conventional solvents such as tetrahydrofuran, acetonitrile and the like.

The acylation reaction can be carried out at a temperature of from -5000 to + 50 ° C. However, it is preferably carried out at or below room temperature and in particular at a temperature from -30 ° C to 0 ° C. It is generally preferred to acylate the compound of formula XVI with an approximately stoichiometric amount of the acylating agent of formula XVII although a slight excess (e.g. 5-25%) of the acylating agent may be used.

It is preferred that the compound of formula XVI be acylated in the form of its N-silyl derivative (using an anhydrous reaction medium). This is conveniently done in situ by simply adding a suitable silylating agent (for example N, O-bistriemethylsilylacetamide) to the solution of compound XVI before the addition of the acylating agent of formula XVII. It is preferred to use about 3 moles of silylating agent per mole of compound XVI although this is not critical. The silyl compound is easily removed after acylation by the addition of water.

The tertiary amines of the formula / RB CH -N 3 -R4 used in the preparation of the compounds of formula I are commercially available or can be readily prepared by methods known in the art.

The invention is further illustrated by the following working method Preparation of starting material Example gel A 'c-cooc H / § u 25 Trim / Å s2 oR2 III Ethyl (Z) -2-methoxyimino-2- (2-tritylaminothiazol-4-yl) acetate (Ina) A mixture of 5.00 g (10.9 mmol) of ethyl (Z) -2-hydroxyimino-2- (2-tritylaminothiazol-4-yl) -acetate (II), 2.04 ml ( 32.8 mmol) of CHBI and 4.54 g of 62.8 mmol) of K 2 CO 3 in 100 ml of dry dimethyl sulfoxide (DMSO) were stirred overnight at room temperature and then poured into 250 ml of water. The precipitate formed was collected by filtration, washed with water and dried to give 5.15 g of the title compound (quantitative yield), m.p. 115 ° C (dec.).

NMR = acDcls ppm 1.32 (an, t), 3.98 (3H, s), 4.30 (za, q), 6.42 (1H, s), 7.2 (1H, ml 7, (IIIH) 455 092 19 Compound IIIb was prepared by the general procedure indicated above but replacing the methyl iodide with ethyl iodide.

Compound R2 Yield (%) M.p. (° C) Liter value (1) M.p. (C) Izza methyl 1oo 11s ° (sana4 ca. 12o ° (sun.) Izïb ethyl of 97-9e ° * * The ester was hydrolyzed without isolation (1) wetrahearQn ¿¿, (197a) 2233 Exemgel BI --- coon ÄH IV Tran onz (Z) -2-methoxyimino-2- (2-tritylaminothiazol-4-yl) acetic acid (IVa) 6.00 g (12.7 mmol) of the ethyl ester IIIa prepared in Example A in 120 ml of ethanol were treated with 12.7 ml of 2N NaOH overnight at room temperature; The reaction mixture was adjusted to pH 8 by adding powdered dry ice and the solvent was evaporated under reduced pressure.

The residue was dissolved in 100 ml of water and the solution was acidified with 1N HCl to pH 2 and then extracted with 3x50 ml of ethyl acetate. The combined extracts were washed with a saturated aqueous solution of sodium chloride, dried and evaporated. The residue was crystallized from ethyl acetate-hexane to give 5.56 g (98% yield) of the title product, m.p. 138-143 ° C (dec.).

NMR = δ ° D 13 δ ppm 3.89 (au, S), 6.52 (1n, s), 7.2 (1sn, S).

Compound IVb was prepared by the general procedure set forth above. 453 092 Compound R2 Yield (%) M.p. (° C, Literature return value (U Sun.) M.p. (C, Sun.) IVa methyl 98 138-143 ca. 140 Ivb ethyl 85 140-145 not reported (1) wetrehed: § 5, (197s) 2233 Example C Benzhydryl-3-hydroxymethyl-7-phenylacetamido-3-cephem-4-carboxylate (VIII) To a stirred suspension of 162.5 ml of phosphate buffer (pH 7) and 20 g of dry wheat bran at At room temperature, g (12.1 mmol) of 7-phenylacetamidocephalosporanic acid sodium salt was added in one portion, the reaction was monitored by HPLC until the hydrolysis was complete (5 hours), the suspension was filtered to remove wheat bran and the filtrate was cooled to 5-10 °. To the cooled solution was added 32 ml of methylene chloride, followed by 24 ml of a 0.5 M solution of diphenyldiazomethane in ethylene chloride, the pH was then adjusted to 3.0 with 28% phosphoric acid.

After one hour, the reaction mixture temperature was allowed to rise to 20 ° C. 56 ml of heptane were added slowly and the resulting crystalline title product was recovered by filtration. The yield of the title product was 3.0 g (50%).

Example D Benzhydryl 7-amino-3-chloromethyl-3-cephem-4-carboxylate (V) To a slurry of 8.3 g (40 mmol) of PCl 5 in 100 ml of CH 2 Cl 2 was added 3.2 g (40 mmol) of pyridine and the mixture was stirred at 20 ° C for 20 minutes. To the mixture was added 5.1 g (10 mmol) of the benzhydryl 3-hydroxymethyl-7-phenylacetamido-3-cephem-4-carboxylate prepared in Example D in a portion with stirring at -40 ° C. The mixture was stirred at 41 ° C for 15 minutes and then allowed to stand at -1 ° C to -1 ° C for 7 minutes. To the cooled solution (-20 ° C) was added 10 ml of propane-1,3-diol and the mixture was allowed to stand at -20 ° C for 16 hours and then for 20 minutes at room temperature with stirring. The resulting solution was washed with 2x20 ml of ice-water and 10 ml of a saturated aqueous solution of sodium chloride, dried over magnesium sulfate and concentrated in vacuo. The gummy residue (12 g) was dissolved in a mixture of CHCl 3 and n-hexane (2: 1) and chromatographed using a silica gel column (200 g) and the same solvent as eluent. Fractions containing the title compound were evaporated in vacuo and the residue was triturated with n-hexane to give the title product (2.1 g; 51%) with a melting point in excess of 110 ° C (dec.). -1 in = vKBr 3400, zsoo, 1785, 1725 cm. 1% 1 cm A uv = Et ° H 265, nm (E max 160). una: ° Ms ° 'de * CDCl 3 3.69 (zu, sa, 4.43 (zu, S), s, os (1n, PPN d, J = 4.5Hz), 5.24 (1H, d, J = 4.5Hz), 6.87 (1H, s), 7.3 (10H, m).

Example E Benzhydryl 3-chloromethyl-7- [XZ) -2-methoxyimino-2- (2-triethylaminothiazol-4-yl) acetamide] -3-cephem-4-carboxylate (VIa) 2.29 g (5, 52 mmol) of the benzhydryl 7-amino-3-chloromethyl-3-cephem-4-carboxylate prepared in Example D in 57 ml of CH 3 CN was treated with 4.09 ml (16.6 mmol) of bis (trimethylsilyl) acetamide ( BSA) 50 minutes at room temperature to obtain a simple solution. To the solution was added an acid chloride solution, which was prepared starting from 2.04 g (4.60 mmol) of (Z) -2-methoxyimino-2- (2-tritylaminothiazol-4-yl) acetic acid (IVa) and 1.15 g (5.52 mmol) PC15 in 20 ml of methylene chloride.

The mixture was stirred for 30 minutes at room temperature, poured into 200 ml of cold water and extracted with 3x100 ml of ethyl acetate. The combined extracts were washed with an aqueous solution of sodium chloride, dried and evaporated.

The remaining syrup (4 g) was chromatographed on a silica gel column (150 g) eluting successively with 10: 1 and 3: 1 mixtures of toluene and ethyl acetate. The fractions containing the desired compound were combined and evaporated to give 2.61 g (68%) of compound VIa as an amorphous powder.

NMR δ GCDCl 3 ppm 3.50 (za, S), 4.02 (sn, S), 4.33 (zu, S), 4.93 (1n, d), 5.87 (1n, q), δ , 65 (1n, S), 6.90 (1H, S), 7.3 (zsn, m).

Example F Benzhydryl 3-iodomethyl-7- [12Z) -2-methoxyimino-2- (2-tritylaminothiazol-4-yl) acetamide] -3-cephem-4-carboxylate (VIIa) A mixture of 1, 50 g (1.79 mmol) of it in eczema; The prepared 3-chloromethyl derivative (VIa) and 1.34 g (8.93 mmol) of NaI in 30 ml of methyl ethyl ketone were stirred for one hour at room temperature. After evaporation of the solvent, the residue was dissolved in 100 ml of ethyl acetate and washed with water, aqueous Na 2 S 2 O 3 and an aqueous solution of sodium chloride, dried and evaporated to give the title compound VIIa (1.47 g, 89%) as an amorphous powder.

NMR = δ cycles ppm 3.55 (za, Aßq), 4.00 (3H, S), 4.25 (zu, 5), 4.97 (1n, a), 5.80 (1H, q), δ , 65 (1n, S), 6.90 (1H, 5), 7.3 (zsn, m).

Example G Benzhydryl-3-chloromethyl-7- [YZ) -2-ethoxyimino-2- (2-tritylaminothiazol-4-yl) acetamide] -3-cephem-4-carboxylate (VIb) To a solution of 1.095 g ( 2.4 mmol) (Z) -2-ethoxyimino-2- (2-tritylaminothiazol-4-yl) acetic acid (IVb) in 20 ml of dichloromethane was added 500 mg of phosphorus pentachloride. After stirring for one hour at room temperature, the mixture was added in one portion to an ice-cold solution of 1.083 g (2.4 mmol) of compound V and 1 ml of BSA in 20 ml of dichloromethane. After stirring for 0.5 h, the reaction mixture was poured into 200 mL of a 10% aqueous solution of sodium bicarbonate and extracted with 100 mL of CHCl 3. The extract was washed with water, dried over magnesium sulfate and evaporated under reduced pressure. The residue was chromatographed on a silica gel column. Elution CHCl 3 gave compound VIb as an amorphous powder in an amount of 1.76 g (86%).

NMR = sCDCIB ppm 1.40 (BH, t, cH 2 cH 3), 3.53 (2H, ABq, 2-cH 2), 4.37 (2H, s, -CH 2 Cl), 4.60 (ZH, q, -CH 2 CH 3 ), 4.90 (1H, d, 6-H), 5.89 (1H, d, 7-H), 6.88 (1h, s, thiazole-H), 6.91 (1H, s, benzhydryl - CH).

Example H H Diphenylmethyl-7-γYZ) -2-ethoxyimino-2- (2-tritylaminothiazol-4-yl) acetamide] -3-iodomethyl-3-cephem-4-carboxylate (VIIb) A mixture of 1.07 g (1 25 mmol) of the compound vib prepared in Example G and 562 mg (2.75 mmol) of NaI in 20 ml of acetone were stirred for 1 hour. The mixture was filtered and the filtrate was poured into water and extracted with ethyl acetate. The organic layer was washed successively with a 5% aqueous solution of Na 2 S 2 O 3, water and a saturated aqueous solution of sodium chloride, dried over magnesium sulfate and evaporated to give 1.04 g (89%) of compound VIIb.

NMR δ CDCl 3 ppm 3.55 (Zh, q, 2-CH 2), 4.27 (2H, s, CH 2 O), .0 2 (1H, d, 6-H), 5.87 (1H, d, 7 -H), 6.68 (1H, s, thiazole ring H), 6.93 (1H, s, benzhydryl-CH). Preparation of Compounds of the Invention Example 1 7- [12Z) -2- (2-Aminothiazol-4-yl) -2-methoxyiminoacetamide] -3- (trimethylammonium) methyl] 3-cephem-4-carboxylate (Ia ) 1 ml (1 mmol) of a 1M solution of trimethylamine in diethyl ether was added to a stirred suspension of 468 mg (0.5 mmol) of benzhydryl-3-iodomethyl-7- [TZ) -2-methoxyimino-2- ( 2-Tritylamino-thiazol-4-yl) -acetamide (3-cephem-4-carboxylate (VIIa) in 30 ml of diethyl ether and the mixture was stirred for 1.5 hours. The precipitated quaternary salt (XVa) was collected by filtration (410 mg; 82% yield) and 3 ml of trifluoroacetic acid (TFA) was added.

This mixture was stirred for 1.5 hours at room temperature and then evaporated to dryness under reduced pressure at a temperature below 20 ° C. The residue was triturated with ether and the precipitated TFA salt was collected by filtration (yield 365 mg), dissolved in a small amount of methanol and chromatographed on a column of 1.8x20 cm 3 of HP-20 resin.

The column was eluted with about 1 liter of water and then with 0.5 ml of a 30% aqueous solution of methanol. The methanolic eluate was evaporated under reduced pressure at a temperature below 40 ° C and the residue was freeze-dried to give the title crude product (yield 129 mg). The ratio of the A2 / A.3 isomers in the crude product was 1: 2, as determined by HPLC. The product was purified by HPLC (Lichrosorb RP-18, 8x300 mm, eluting with 1 / 100M NH 4 H 2 PO 4 (pH 7.2): CH 3 OH = 85:15). The HPLC eluate was chromatographed on a single column of HP-20 (1.8x15 cm 3) to remove the inorganic salt. The column was eluted with 0.5 liters of water and then with 0.5 liters of a 1% aqueous solution of methanol. The methanolic eluate was evaporated under reduced pressure at a temperature below 40 ° C and the residue was lyophilized to give the title compound (Ia) as an amorphous powder.

Yield 75 mg (33%, calculated on VIIa). The product gradually decomposed above 160 ° C. Estimated purity 80%. 453 092 Example 2 7 - [(Z) -2- (2-aminothiazol-4-yl) -2-methoxyiminoacetamide] -3- [N, N-dimethyl-N- (2-hydroxyethyl) ammonium] methyl-3 cephem-4-carboxylate (Ib) The general procedure of Example 1 was repeated except that the trimethylamine used therein was replaced with an equimolar amount of N, N-dimethylethanolamine. The crude product had an .A2: .A3 ratio of 1: 2. After purification, the title compound was obtained in a yield of 17% and decomposed at a temperature above 160 ° C. Estimated purity 90%.

Example 3 7- [YZ) -2- (2-Aminothiazol-4-yl) -2-methoxyiminoacetamide] -3- (N, N-dimethyl-N-allylammonium) methyl 3-cephem-4-carboxylate (Ic) The general procedure of Example 1 was repeated except that the trimethylamine used therein was replaced with an equimolar amount of N, N-dimethylallylamine. The crude product had an A2: A3 ratio of 1: 4.5. After purification, the title compound was obtained in 14% yield and decomposed at a temperature above 150 ° C. Estimated purity 80%.

Example 4 7 - [(Z) -2- (2-aminothiazol-4-yl) -2-methoxyiminoacetamide] -3- [N, N-dimethyl-N- (3-pyridylmethyl) ammonium] methyl-3-c five-4-carboxylate (Id) The general procedure of Example 1 was repeated except that the trimethylamine used therein was replaced with an equimolar amount of 3- (dimethylaminomethyl) pyridine. The crude product had an A2: A3 ratio of 1: 4.3. After purification, the title compound was obtained in a yield of 17% and decomposed at a temperature above 170 ° C.

Estimated purity 75%. Example 5 7- [12Z) -2- (2-Aminothiazol-4-yl) -2-methoxyiminoacetamide] -3- [N, N-dimethyl-N- (2-dimethylaminoethyl) ammonium] methyl -3- cephem-4-carboxylate (Ie) The general procedure of Example 1 was repeated except that the trimethylamine used therein was replaced with an equimolar amount of 1,2-bis (dimethylamino) ethane. The crude product had a 1: 1 ratio of 1: 1. After purification, the title compound was obtained in a yield of 14% and decomposed at a temperature above 150 ° C. Estimated purity 65%.

Example 6 7- [YZ) -2- (2-Aminothiazol-4-yl) -2-methoxyiminoacetamide] -3- (N, N-dimethyl-N-ethylammonium) methyl 3-cephem-4-carboxylate ( The general procedure of Example 1 was repeated except that the trimethylamine used therein was replaced with an equimolar amount of N, N-dimethylethylamine. The crude product had an A2: ¿S3 ratio of 1: 1. After purification, the title compound was obtained in a yield of 15% and decomposed at a temperature above 150 ° C. Estimated purity 77%.

Example 7 7 - [(Z) -2- (2-aminothiazol-4-yl) -2-methoxyiminoacetamide] -3- (N, N-diethyl-N-methylammonium) methyl 3-cephem-4-carboxylate (Ig) The general procedure of Example 1 was repeated except that the trimethylamine used therein was replaced with an equimolar amount of diethylmethylamine. The crude product had a §2: A 3 ratio of 1: 1. After purification, the title compound was obtained in a yield of 10% and decomposed at a temperature above 150 ° C. Estimated purity 65%. Example 8 7 - [(Z) -2- (2-Aminothiazol-4-yl) -2- (ethoxyiminoacetamide] -3- (triethylammonium) methyl 3-olefin-4-carbonylate (Ih) The general procedure of Example 1 was repeated except that benzhydryl-3-iodomethyl-7- [YZ) -2-methoxy-imino-2- (2-tritylaminothiazol-4-yl) acetamido] -3-cephem-4-carboxylate (VIIa) was replaced with an equimolar amount of the corresponding ethoxyimino compound (VIIb) (prepared according to Example H). The crude product had an A2; LS3 ratio of 3: 1. After purification, the title compound was obtained in 3% yield and decomposed at a temperature above 150 ° C. Estimated purity 70%.

Spectral data for the compounds according to examples 12-8 (a) Infrared spectra (KBr) All products showed similar infrared spectra: 1770-1775 cm * (e-: Lactam c = o), 1660: m4 (conm, 1610 cm * (cooï. ( b) Ultraviolet spectra (1/15 M phosphate buffer; pH 7) All products except the product according to Example 4 have similar spectra: 235 nM (a 15700-16400), 257 nM (6 15400-16000). the following values: 235 nM (E 17600), 255 nM (s 18600, sk), 260 nM (s 19000) 266 nM (e 17900, sk) 455 092 28 (c) NMR sectra (D20) Fönaüng Ka fi sk: & k $ quh gym ng gym) enl. + 6-H 7-H example N -qla 0-01 (1H, d, (1H-, d, iiazol-H (s) (small) 4-5Hz; 4-snz ) mms) other 2 3.20 (3H) 4.10 5.46 5.90 7.10 3.25 (3H) 3 3.08 (3H) 4.10 5.46 7.10 Saw-6.4 3.15 (an) åkåšfn '4 3.04 (3H) 4.09 5.45 5.94 7.06 4.75 (2H, s, PYrf-'fí) 3.21 (an) 7.66 (nad-d. 8 & 4Hz, PyrrH) 8.12 (1H, d, 8Hz, Pyr-H) 8.74 (2H, s, Pyf- * fl 3.18 (3H) 4.10 5.45 5.90 7.10 2.50 ( 6H, s, 3.25 (3H) N (CH 3) 2) δ 3.10 (3H) 4.10 5.45 5.90 7.10 1.48 (3H, t , 3.15 (3H) 7Hz, CH2q§3) δ 3.05 (3H) 4.10 5.44 5.95 7.10 1.45 (6H, t, 7Hz, CH2Q§3) δ 3.25 (9H ) 7 ”5,50 5,97 7,10 1,45 (3H, t, 7Hz¿H2qh) It has been shown that both the A2: .3 the quaternary salt (XV) is allowed to stand for a short period of time and is re-filtered through the bed of the originally recovered compound (XV), especially if this re-filtration step is repeated several times. The yield can thus be increased slightly if a small amount of the original tertiary amine reactant is added to the filtrate. This is shown in the following example, which is a modification of Example 1. 453 092 29 Example 9 7 - [(Z) -2- (2-aminothiazol-4-yl) -2-methoxyiminoacetamide] -3- (trimethylammonium) methyl- 3-Cephem-4-carboxylate (Ia) ml of a 1M solution of trimethylamine ether was added in one portion to a stirred solution of 4.68 g (5 mmol) of the iodide compound VIIa in 500 ml of ether. The mixture was stirred for minutes and the precipitated quaternary salt (XVa) was collected by filtration and washed with a small amount of ether. The filtrate and washings were combined and allowed to stand at room temperature for a further 10 minutes and the second precipitate which separated was collected by filtration on the same funnel where the first precipitate was kept as a bed.

The filtrate was filtered three times in the same manner at 10 minute intervals for obtained. of 3.58 g (72% starting from VIIa) of the quaternary salt (XVa). To the final filtrate was added 2 ml of a 1M solution of trimethylamine in ether, and the same filtration operation was repeated 3 more times at 10 minute intervals to obtain 0.56 g (11%) of compound Xva. To the obtained filtrate was added another 1 ml of a 1M trimethylamine solution, and the refiltration operation was repeated twice at minute intervals to obtain a third batch of compound Xva (0.134 g; 2.7%). The total yield of compound XVa was 4.27 g (86%).

A mixture of 4.20 g (4.22 mmol) of compound XVa, 1 ml of anisole and 40 ml of TFA was stirred for 1.5 hours at room temperature. The mixture was evaporated under reduced pressure at a temperature below 20 ° C and the dark residue was triturated with 300 ml of isopropyl ether to precipitate 3.50 g of the TFA salt, which was collected by filtration and dried under reduced pressure. The TFA salt was dissolved in 50 ml of methanol, treated with a small amount of charcoal and filtered. The filtrate was concentrated under reduced pressure, and ml of a 1M solution of sodium 2-ethylhexanoate in ethyl acetate was added to the concentrate. The mixture was diluted with 300 ml of ethyl acetate to precipitate 2.36 g of the title compound as a crude product (estimated purity 50%), which was collected by filtration, washed with a small amount of ethyl acetate and dried. The ratio of Az-isomer to Å3-isomer of the crude product was 1: 4 (HPLC, Lichrosorb RP-18; mobile phase, 1 / 100M ammonium phosphate buffer, pH 7 - cH3oH, 90:10; Retention time, A2-isomer 6 ' 54 ", ¿_ \. 3-isomer 8 '29").

The crude product (2.36 g) was dissolved in a small amount of water and purified using HPLC (Waters Associates, System 500, PrepPAK 50O / C18; mobile phase, 7% methanol). The HPLC eluate containing the product was concentrated under reduced pressure at a temperature below .35 ° C and the concentrate was lyophilized to give the title compound. Yield 959 mg (42%, based on iodide VIIa). Amorphous powder. Estimated purity 80% (according to HPLC). The ratio of Az-isomer to 1A3-isomer was 1:17. The compound gradually decomposed at a temperature above 160 ° C. fi '

Claims (9)

10 15 20 25 30 35 453 092 31 PATENT REQUIREMENTS 1. Compounds of formula 1 /: § LW? R HN S NH --- r el: I P-li ,, / c? wherein R 1 is hydrogen, R 2 and R 3 are each independently methyl or ethyl and R 4 is methyl, ethyl, allyl, 2-hydroxyethyl, 2- (dimethylamino) ethyl or pyridylmethyl and non-toxic pharmaceuticals. culturally acceptable southern addition salts and solvates thereof. Compounds of formula I according to claim 1, characterized in that R 1 is hydrogen, R 2 is methyl, R 3 is methyl or ethyl and R 4 is methyl, ethyl, 2-hydroxyethyl, 2- (dimethylamino) ethyl, allyl or pyridylmethyl, and non-toxic pharmaceutically acceptable acid addition salts and solvates thereof. 3. The compounds 7- [2Z) -2- (2-aminothiazol-4-yl) -2-methoxyiminoacetamide] -3- (trimethylammonium) -methyl-3-cephem-4-carboxylate, 7- [1Z) - 2- (2-aminothiazol-4-yl) -2-methoxyiminoacetamide Q7-3- [W, N-dimethyl-N- (2-hydroxyethyl) ammonium methyl-3-cephem-4-carboxylate and 7 [TZ) -2- ( 2-aminothiazol-4-yl) -2-methoxyiminoacetamide (7-N-dimethyl-N-allylammonium) methyl 3-cephem-4-carboxylate according to claim 1 and non-toxic, pharmaceutically acceptable acid addition salts and solvate thereof. 4. An antibacterial composition, characterized in that it comprises an antibacterially effective amount of a compound according to claim 1 and an inert pharmaceutical carrier. An antibacterial composition according to claim 4 in unit dosage form, characterized in that it comprises 50-1500 mg of a compound according to claim 1 and an inert pharmaceutical carrier. 10 15 20 25 30 35 453 092 n An antibacterial composition according to claim 4 or 5, characterized in that it comprises the compound 7 [] Z) -2- (2-aminothiazol-4-yl) -2-methoxyiminoacetamide g7- 3- (trimethylammonium) methyl- 3-cephem-4-carboxylate according to claim 1 or non-toxic pharmaceutically acceptable acid addition salt or solvate thereof. 7. Process for the preparation of the compounds according to nu E s N;, »I If a” _ mf: - ï- “a co - RÅ N C l \ ll. N \ 2 OR H28 S. each is independently methyl or claim 1 of the formula wherein R 2 and R 3 are ethyl and R 4 is methyl, ethyl, allyl, 2-hydroxyethyl, 2- (dimethylamino) ethyl or pyridylmethyl and non-toxic pharmaceutically acceptable salts and solvates thereof, characterized in that (A) is reacting a compound of the formula in ß m. S u, Jif / wherein R2 has the meaning given above, B1 is a conventional carboxyl-protecting group and B2 is a conventional amino-protecting group, with a tertiary amine with the formula f: cn3-u \ R4 5 10 15 20 25 30 35 ~ uzu S \\\ OR? 453 092 33 wherein R3 and R4 have the meanings given above, for the preparation of a compound of the formula ä Miri / S 3 R xv N (91 / cn -rlv-cn co 4 g 2 3 R and that all protecting groups are subsequently removed in a conventional manner and / or if desired, the compound of formula I converts to a non-toxic pharmaceutically acceptable salt or solvate thereof or to B) acylate a compound of formula. xvI - islands-N 1 / 'C Hi - N- ~ CH 3 coon * å * 1 ar hydrogen or a conventional carboxyl protecting group and R 3 and R 4 have or an N-silyl derivative thereof, wherein B is as defined above, with an acylating derivative of an acid of formula c-Äcoon N 132m / qlšu \ onz xvn wherein B2 is a conventional amino protecting group R2 has the meaning given above, for the preparation of a compound of formula 10 15 20 25 30 35 5 aämlïšší 453 092 34 S "rf NO / É / m; and that all protecting groups are then removed in a conventional manner and / or, if desired, the compound of formula I is converted into a non-toxic pharmaceutically acceptable salt or solvate thereof. A process according to claim 7 for the preparation of compounds of the formula O 4 - R 4 is hydrogen, R 2 is methyl, R 3 is methyl or ethyl. and R 4 is methyl, ethyl, 2-hydroxyethyl, 2- (dimethylamino) ethyl, allyl or pyridylmethyl, or a non-toxic pharmaceutically acceptable acid addition salt or solvate thereof characterized by reacting a mixture of benzhydry1-7-amino- 3-Chloromethyl-3-cephem-4-carboxylate and bis- (trimethylsilyl) acetamide with an acid chloride of (Z) -2-methoxyimino-2- (2-tritylaminothiazol-4-yl) acetic acid in an organic solvent to obtain of the corresponding 3-chloromethyl-3-cephem derivative, then reacting the 3-chloromethyl-3-cephem derivative with an alkali metal iodide salt in an organic solvent to obtain the corresponding 3-iodomethyl-3-cephem derivative, then reacting The 3-iodomethyl-3-cephem derivative in an organic solvent with trimethylamine or N, N-dimethylethanolamine or N, N-dimethylallylamine or 3- (dimethyl laminomethyl) pyridine or 1,2-bis (dimethylamino) ethane or N, N-dimethylethylamine or dimethylethylamine to obtain the corresponding quaternary salt and finally blocking said quaternary salt for the obtained of 7¿ÅZ) - 2- (2-aminothiazol-4-yl) -2-methoxyiminoacetamide (3-trimethylammonium) methyl 3-cephem-4-carboxylate (Ia) or 7-ÅZ) -2- (2-aminothiazol-4-yl) - 5-Ethoxyiminoacetamido] -3- [N, N-dimethyl-N- (2-hydroxyethyl) ammonium] methyl 3-cephem-4-carboxylate (Ib) or 7- [1Z) -2- (2-aminothiazole) 4-yl) -2-methoxyiminoacetamide] -3- [N, N-dimethyl-N-allylammonium methyl 3-cephem-4-carboxylate (Ie) or 7- [YZ) -2- (2-aminothiazol-4-yl) ) -2-methoxyiminoacetamide] -3- [N, N-dimethyl-N- (3-pyridylmethyl) ammonium] methyl 3-cephem-4-carboxylate (Id) or 7- [1Z) -2- (2-aminothiazole -4-yl) -2-methoxyiminoacetamide [3- [N, N-dimethyl-N- (2-dimethylaminoethyl) ammonium] methyl 3-cephem-4-carboxylate (Ie) or 7- [η 2) -2 - (2-Aminothiazol-4-yl) -2-methoxyiminoacetamide] -3- [N, N-dimethyl-N-ethylammonium fl methyl-3-cephem-4-carboxylate (If) or 7- [XZ) -2- (Sw-thiaziazol-4-yl) -2-methoxyiminoacetamide] -3- [N, N-dimethyl-N-methylammonium methyl-3-cephem-4-carboxylate (Iq) And / or, if desired, converting said compounds (Ia) to (Ig) into non-toxic pharmaceutically acceptable acid addition salts or solvates thereof. A process according to claim 7 for the preparation of the compound 7-112) -2- (2-aminothiazol-4-yl) -2-ethoxyiminoacetamide] -3- (trimethylammonium) methyl-3-cephem-4-carboxy lat (Ib), characterized reactes a mixture of benzhydryl 7-amino-3-chloromethyl-3-cephem-4-carboxylate and bis (trimethylsilyl) acetamide with an acid chloride of (Z) -2-ethoxyimino-2 - (2-tritylamino-thiazol-4-yl) -acetic acid in an organic solvent to give the corresponding 3-chloromethyl-3-cephem derivative, then the 3-chloro-methyl-3-cephem derivative reacts with an alkali metal iodide salt to obtain of the corresponding 3-iodomethyl-3-cephem derivative, then the 3-iodomethyl-3-cephem derivative reacts with trimethylamine in an organic solvent to give the corresponding quaternary salt and finally unblocks the quaternary salt to obtain hence, that one of the compound (Ih).